Distemper serum and method of use



35. the preventative agent '55 admission and minis July 24, 1934 UNITED STATES ATENT OFFICE i ms'ranmin asserts: mmrop or van Georg e Watson Little, New York, N. in

No Drawing. Application December 5 im, Serial No. 5.15s

a mum. (on. 1014s;

This invention relates to the prevention and the treatment or distemper. The inventiommore' particularly, relates to serum preparation and use and has 'ior an object the provision or an im- 8 proved serium and method 01' use.

The invention aims to provide an improved serum and a method or preparing it, It further aims to provide a method 01' treating dogs to produces. lasting immunization against distemper IQ and to provide treatment for dogs ailected with v distemper for their immunization and cure.

' A method used heretofore for the prevention or distemper involves the. simultaneous injection of a serum on one side of a dog and an antigen '15 on the other. This method of treatment producesa temporary immunity against distemper. An-

other method heretoi'ore used for the prevention 01' distemper involves the use of vaccine and living virus oi Carre. The vaccine is which is followed by the injection etliving virusi r method a solid or lasting immunity In this latte is accomplished by using a. vaccine containing a preventative agent sumciently strong to prevent vthe disease when the living virus is injected. One

disadvantage in such treatment is attributed to formalin. which s at the point 01 injection.

A method for the preparation oi serum hereto .tore used is carried out by injecting virus subcutaneously or intra-musculariy into immune .40 animals. Such method of injection has been found to be objectionable for the reason'that the strength of injected agent is lost in the process of absorption. Attempts have been made heretofore to inject virus fluid or suspension intravenously but such attempts have only resulted in thrombosis or other objectionable results which have prevented method. 1

The invention contemplates the preparation or any practical application oi that I serum of such concentration that it will hold the According to a method of- -iivlng virus at Carre.

the invention, living virus is injected intravenously into an'immune dog to produce serum. In carrying out such intravenous injection the rate of temperature oi the living virus injectedac I living virus causes fluid is so regulated that no appreciable injury resultsto the dog. Advantages over the heretoi'ore used methods-oiinjection are manifested in elimination of losses due to absorption and in the ability to produce a concentrated serum.

In the treatment of dogs; the invention contemplates the simultaneous injection of living virus of Carr and serum for immunization against dieof a dog having distemper Puppies as rise in temperature. Thus puppies can be protected before the, susceptible age, which is not true 01' methods now used.

In preparing the serum to be used in practic- 35 ing the inventions. living virus sections and placed between-the layers of sterile gauze. Pressure is applied by twisting the gauze and the spleen therein is then thoroughly macerated; Sterile normal saltsolution is added in an amount 'sumcient to make a 20% suspension of spleen tissues In other words;- a spleen weighing 20 grams' for use, at which time it bath and centrifuged for twenty to thirty minutes in preparation for use.

.This living virus suspension isinjected into the 1 jugular vein 01' an immune dog.

The immunity rig .painted with iodine.

of a dog must be carefully established by running a series of injections of the living virus .over a period of two or three weeks to determine positively whether or not the dog is immune.

The immune dog is prepared for the injection of a living virus suspension by-clipping and shaving the area known as the jugular furrow of the neck. The skin is then thoroughly cleansed 'and By stoppin the flow of bloodin the jugular notch the vein is distended and its location becomes clearly discernible. A sterile hypodermic needle, which is preferably of 21 gauge, is then thrust through the skin into the jugular vein to the pressure applied. When blood flows back through the hypodermic needle a syringe containing cc. of 20% suspension of the living virus, at body heat,'is coupled to the hypodermic needle. The suspension is then injected very slowly into the jugular vein for a period of one minute or longer. When time is taken to inject.

. the suspension of the warm living virus sumciently slowly, there is no danger of causing a reaction manifested by fainting and loss of consciousness. If the centrifuged solution is injected rapidly and below the body temperature of the dog, a reaction resulting in fainting or loss of consciousness is almost certain to occur.

a as the jugular furrow.

in the 'skin and an incision one-v bottle prevents the Danger of throbosis (occlusion of the arteries with a foreign substancel is prevented by the above regulation of the admission of the suspended fiuid and its temperature.

Injections of the living virus in the jugular vein may be made for three succeeding days in each week for three weeks, followed by bleeding seven days after the last injection. An injection may also be made in the jugular vein for two days which is followed by bleeding seven days after the last injection.

The dogs are prepared for bleeding by shaving and cleansing the skin area on the neck known A solution of 2% butyn is injected quarter inch long is made. A large gauge needle is inserted through the wound. The jugular vein is filled up by exerting pressure with the thumb in the jugular notch so that it stands out like a cord. The needle is thrust quickly through the wall of the vein, whereupon the blood flows in a steady stream into a sterile bottle. A quantity of a weak solution of sodium citrate in the blood from clotting when it is agitated continuously. Bleeding proceeds until the bottle is filled. A dog the size of a German shepherd will yield from 500 to 700 cc. of blood without any visible signs of weakness.

The blood is. refrigerated and taken to the laboratory whereupon the serum is made, concentrated, preserved and bottled as follows:

1. The blood is centrifuged to remove the fibrin whereupon the plasma remaining is. about 50% of the total volume of the blood.

2. The plasma is diluted 5.0% with water and precipitated with ammonium sulphate up to 30% saturation. It is then filtered and washed with a quantity of water-equal to half the quantity oflthe plasma. It is then filtered again. Then both filtrates are precipitated with ammonium sulphate up to 50% saturation.

3. It is then filtered, pressed and dialyzed.

4. The filtrate is preserved with a solution of Merthiolate l-l0,000 cc. to the liter and 1% of sodium chloride added whereupon it is filtered through a Berkefeld filter and bottled in sterile bottles containing 4 cc. each. The resulting conwhich stands out plainly due" centrated serum is a pale transparent liquid five times the strength of normal homologous serum.

In immunizing a puppy against distemper it is first determined whether the animal is in a normal healthy condition. The temperature and a microscopic test of the feces are taken to determine the presence or absence of'worms "(intestinal parasites). Aninquiry into the history of the dog is made in order to determine whether he has ever had distemper or a cold. The serum is' then removed from the bottle by inserting the hypodermic needle through the rubber diaphragm of the cork and drawn into the barrel of a sterile I hypodermic needle. The skin of the shoulder or neck areais clipped, shaved, cleansed and painted with iodine. By a quick thrust of the hypodermic needle the serum (4 cc.) is injected underneath the skin.

Living virus is obtained from dried virus which is placed in a small rubber stoppered bottle containing a sterile hypodermic syringe and needle. One quarter of 1 cc. of normal salt solution is drawn into the syringe which is injected into the bottle through the rubber stopper and mixed with the dried virus whereupon 'it is drawn back into the syringe again. An area of skin in the shoulder or neck region is then clipped, shaved, cleansed and painted with iodine. The living virus thus obtained is injected between the layers of the skin (intradermally) by grasping the skin with the left forefinger and thumb and piercing the layers of the skin with the hypodermic needle, care being taken to direct the needle parallel to the level of the left forefinger and thumb. Care is .also taken not to pierce the skin too deeply, otherwise the virus solution will be injected beneath the skin layers.

Of course, great care should be taken that all dogs be examined in order to ascertain whether they are immune, have been inoculated previously, or exposed to distemper. These .factors cannot always be determined.

For example, a puppy ten weeks old may have such a history that it is difiicult to know whether or not it is or has been affected with distemper. Under such circumstances it is advisable to inject 4 cc. of serum alone, subcutaneously, when there is no elevation of temperature or other symptoms present. If there are no symptoms of the disease and temperature is normal, in from two to five 'days a second injection of serum and simultaneously living virus are administered. In case there is a doubt that the puppy is in the incubation stage, this procedure will suiliciently protect it. It-proves that the puppy is susceptible or not for in a few days, if efiected, some signs will appear such as a slight nasal discharge indicating that the puppy was affected by the disease, or, if not afiected,'it will remain normal. The outstanding fact being that the puppy passes through only a mild form of the disease without any danger or necessity of further treatment.

Thus in the method of treatment with a serum of the invention, dogs having distemper in the incubative stage, before any signs are evident,

1,987,478 dose of 4 cc. asoften as every forty-eight hours.

A number oi' experiments have been made to determine relative serum concentration. It has been found that serum obtained from an immune dog that had been intravenously injected with living virus in periods of three succeeding days for three weeks and blood withdrawn seven days alter the last virus injection was 0! such character that a concentrated serum could. be produced therefrom. v

A concentrated serum, as referred to herein, is intended to mean a serum 01' such character that it is capable 01 holding the living virus of Carr.

During the course of experimentation it was determined that approx-imately'i cc. of the concentrated serum comprised the minimum lethal dose. The minimum dose of concentrated serum 7 is determined, in part, by the smallest amount that is required to hold the living virus, thereby preventing it from causing distemper.

I claim:

1. In a process ior preparing distemper serum, the step which comprises injecting living virus 01' Carr of spleenic origin intravenously into an immune dog, withdrawing bloodriromsaid'rlog and extracting serum therei'rom.

2. In a process ior preparing distemper the step which comprises injecting a twenty per cent suspension oiliving virus oi Carr intravenously into an immune dog, withdrawing blood from said dog and extracting serum therefrom. I

"sc -ism! "read scr m; page}, line 29.

3. Ina process for preparing distemper serum, 7 the improvement comprising injecting living virus oi Carr of spleenic origin into the jugular vein of an immune dog, withdrawing blood from said dog and extracting serum therefrom.

4. A distemper preparationior the treatment ,0! dogs comprising a serum obtained by injecting living virus of Carr oi spleenic origin intra venously into an immune dog, withdrawing blood from the dog and preparing a concentrated serum therefrom adaptable tor the simultaneous injection withliving virus of Canto produce distemper immimization.

5. A distemper preparation for the treatment oi dogs comprising a serum obtained by iuJecting. living virus or Carr 'oi spleenic origin intravenously into a dog for three consecutive days oi each week ior three weeks, bleeding the dog approximately seven days after the last virus iniection, and extracting a concentrated serum from the blood.

6. A distemper preparation !or the treatment of dogs comprising a serum obtained by iniecting living virus oi. Carr oi spleenic origin intravenously into a dog in two applications. preterahly on succeeding days. and approximately seven days later removing blood from said dog iromwhich the serum is produced.

amonqn wa'rson Watson ,7 His hereby certified ihst error sppesrs 'in the printed speeiiies'tion oi the 1 above numbered palem requiring correction as follows: Page" i, line 5,' for for "throbosis"- read thrombosis; and I page 3; "line'9l,.clai|ii"5, .sttikebut the words -"oi' spleenlc origin"; and that .the said Letters Patent should-be read with these correctionstherein thst the f1 same may conform to the record of the 7 Sigaedjand sealed this 11th day of September, A. D. i934.

case iu'the Patent-Office.

. iiesl ie Fraser 5 omits-mt o titans.

1,987,478 dose of 4 cc. asoften as every forty-eight hours.

A number oi' experiments have been made to determine relative serum concentration. It has been found that serum obtained from an immune dog that had been intravenously injected with living virus in periods of three succeeding days for three weeks and blood withdrawn seven days alter the last virus injection was 0! such character that a concentrated serum could. be produced therefrom. v

A concentrated serum, as referred to herein, is intended to mean a serum 01' such character that it is capable 01 holding the living virus of Carr.

During the course of experimentation it was determined that approx-imately'i cc. of the concentrated serum comprised the minimum lethal dose. The minimum dose of concentrated serum 7 is determined, in part, by the smallest amount that is required to hold the living virus, thereby preventing it from causing distemper.

I claim:

1. In a process ior preparing distemper serum, the step which comprises injecting living virus 01' Carr of spleenic origin intravenously into an immune dog, withdrawing bloodriromsaid'rlog and extracting serum therei'rom.

2. In a process ior preparing distemper the step which comprises injecting a twenty per cent suspension oiliving virus oi Carr intravenously into an immune dog, withdrawing blood from said dog and extracting serum therefrom. I

"sc -ism! "read scr m; page}, line 29.

3. Ina process for preparing distemper serum, 7 the improvement comprising injecting living virus oi Carr of spleenic origin into the jugular vein of an immune dog, withdrawing blood from said dog and extracting serum therefrom.

4. A distemper preparationior the treatment ,0! dogs comprising a serum obtained by injecting living virus of Carr oi spleenic origin intra venously into an immune dog, withdrawing blood from the dog and preparing a concentrated serum therefrom adaptable tor the simultaneous injection withliving virus of Canto produce distemper immimization.

5. A distemper preparation for the treatment oi dogs comprising a serum obtained by iuJecting. living virus or Carr 'oi spleenic origin intravenously into a dog for three consecutive days oi each week ior three weeks, bleeding the dog approximately seven days after the last virus iniection, and extracting a concentrated serum from the blood.

6. A distemper preparation !or the treatment of dogs comprising a serum obtained by iniecting living virus oi. Carr oi spleenic origin intravenously into a dog in two applications. preterahly on succeeding days. and approximately seven days later removing blood from said dog iromwhich the serum is produced.

amonqn wa'rson Watson ,7 His hereby certified ihst error sppesrs 'in the printed speeiiies'tion oi the 1 above numbered palem requiring correction as follows: Page" i, line 5,' for for "throbosis"- read thrombosis; and I page 3; "line'9l,.clai|ii"5, .sttikebut the words -"oi' spleenlc origin"; and that .the said Letters Patent should-be read with these correctionstherein thst the f1 same may conform to the record of the 7 Sigaedjand sealed this 11th day of September, A. D. i934.

case iu'the Patent-Office.

. iiesl ie Fraser 5 omits-mt o titans. 

